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Retrospective species identification of microsporidian spores in diarrheic fecal samples from human immunodeficiency virus/AIDS patients by multiplexed fluorescence in situ hybridization.
J Clin Microbiol. 2007 Apr; 45(4):1255-60.JC

Abstract

In order to assess the applicability of multiplexed fluorescence in situ hybridization (FISH) assay for the clinical setting, we conducted retrospective analysis of 110 formalin-stored diarrheic stool samples from human immunodeficiency virus (HIV)/AIDS patients with intestinal microsporidiosis collected between 1992 and 2003. The multiplexed FISH assay identified microsporidian spores in 94 of 110 (85.5%) samples: 49 (52.1%) were positive for Enterocytozoon bieneusi, 43 (45.8%) were positive for Encephalitozoon intestinalis, 2 (2.1%) were positive for Encephalitozoon hellem, and 9 samples (9.6%) contained both E. bieneusi and E. intestinalis spores. Quantitative spore counts per ml of stool yielded concentration values from 3.5 x 10(3) to 4.4 x 10(5) for E. bieneusi (mean, 8.8 x 10(4)/ml), 2.3 x 10(2) to 7.8 x 10(4) (mean, 1.5 x 10(4)/ml) for E. intestinalis, and 1.8 x 10(2) to 3.6 x 10(2) for E. hellem (mean, 2.7 x 10(2)/ml). Identification of microsporidian spores by multiplex FISH assay was more sensitive than both Chromotrope-2R and CalcoFluor White M2R stains; 85.5% versus 72.7 and 70.9%, respectively. The study demonstrated that microsporidian coinfection in HIV/AIDS patients with intestinal microsporidiosis is not uncommon and that formalin-stored fecal samples older than 10 years may not be suitable for retrospective analysis by techniques targeting rRNA. Multiplexed FISH assay is a reliable, quantitative fluorescence microscopy method for the simultaneous identification of E. bieneusi, E. intestinalis, and E. hellem, as well as Encephalitozoon cuniculi, spores in fecal samples and is a useful tool for assessing spore shedding intensity in intestinal microsporidiosis. The method can be used for epidemiological investigations and applied in clinical settings.

Authors+Show Affiliations

Department of Environmental Health Sciences, Division of Environmental Health Engineering, Johns Hopkins Bloomberg School of Public Health, 615 N. Wolfe Street, Baltimore, MD 21205, USA. tgraczyk@jhsph.eduNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

17287331

Citation

Graczyk, Thaddeus K., et al. "Retrospective Species Identification of Microsporidian Spores in Diarrheic Fecal Samples From Human Immunodeficiency virus/AIDS Patients By Multiplexed Fluorescence in Situ Hybridization." Journal of Clinical Microbiology, vol. 45, no. 4, 2007, pp. 1255-60.
Graczyk TK, Johansson MA, Tamang L, et al. Retrospective species identification of microsporidian spores in diarrheic fecal samples from human immunodeficiency virus/AIDS patients by multiplexed fluorescence in situ hybridization. J Clin Microbiol. 2007;45(4):1255-60.
Graczyk, T. K., Johansson, M. A., Tamang, L., Visvesvara, G. S., Moura, L. S., DaSilva, A. J., Girouard, A. S., & Matos, O. (2007). Retrospective species identification of microsporidian spores in diarrheic fecal samples from human immunodeficiency virus/AIDS patients by multiplexed fluorescence in situ hybridization. Journal of Clinical Microbiology, 45(4), 1255-60.
Graczyk TK, et al. Retrospective Species Identification of Microsporidian Spores in Diarrheic Fecal Samples From Human Immunodeficiency virus/AIDS Patients By Multiplexed Fluorescence in Situ Hybridization. J Clin Microbiol. 2007;45(4):1255-60. PubMed PMID: 17287331.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Retrospective species identification of microsporidian spores in diarrheic fecal samples from human immunodeficiency virus/AIDS patients by multiplexed fluorescence in situ hybridization. AU - Graczyk,Thaddeus K, AU - Johansson,Michael A, AU - Tamang,Leena, AU - Visvesvara,Govinda S, AU - Moura,Laci S, AU - DaSilva,Alexandre J, AU - Girouard,Autumn S, AU - Matos,Olga, Y1 - 2007/02/07/ PY - 2007/2/9/pubmed PY - 2007/10/12/medline PY - 2007/2/9/entrez SP - 1255 EP - 60 JF - Journal of clinical microbiology JO - J Clin Microbiol VL - 45 IS - 4 N2 - In order to assess the applicability of multiplexed fluorescence in situ hybridization (FISH) assay for the clinical setting, we conducted retrospective analysis of 110 formalin-stored diarrheic stool samples from human immunodeficiency virus (HIV)/AIDS patients with intestinal microsporidiosis collected between 1992 and 2003. The multiplexed FISH assay identified microsporidian spores in 94 of 110 (85.5%) samples: 49 (52.1%) were positive for Enterocytozoon bieneusi, 43 (45.8%) were positive for Encephalitozoon intestinalis, 2 (2.1%) were positive for Encephalitozoon hellem, and 9 samples (9.6%) contained both E. bieneusi and E. intestinalis spores. Quantitative spore counts per ml of stool yielded concentration values from 3.5 x 10(3) to 4.4 x 10(5) for E. bieneusi (mean, 8.8 x 10(4)/ml), 2.3 x 10(2) to 7.8 x 10(4) (mean, 1.5 x 10(4)/ml) for E. intestinalis, and 1.8 x 10(2) to 3.6 x 10(2) for E. hellem (mean, 2.7 x 10(2)/ml). Identification of microsporidian spores by multiplex FISH assay was more sensitive than both Chromotrope-2R and CalcoFluor White M2R stains; 85.5% versus 72.7 and 70.9%, respectively. The study demonstrated that microsporidian coinfection in HIV/AIDS patients with intestinal microsporidiosis is not uncommon and that formalin-stored fecal samples older than 10 years may not be suitable for retrospective analysis by techniques targeting rRNA. Multiplexed FISH assay is a reliable, quantitative fluorescence microscopy method for the simultaneous identification of E. bieneusi, E. intestinalis, and E. hellem, as well as Encephalitozoon cuniculi, spores in fecal samples and is a useful tool for assessing spore shedding intensity in intestinal microsporidiosis. The method can be used for epidemiological investigations and applied in clinical settings. SN - 0095-1137 UR - https://www.unboundmedicine.com/medline/citation/17287331/Retrospective_species_identification_of_microsporidian_spores_in_diarrheic_fecal_samples_from_human_immunodeficiency_virus/AIDS_patients_by_multiplexed_fluorescence_in_situ_hybridization_ L2 - http://jcm.asm.org/cgi/pmidlookup?view=long&pmid=17287331 DB - PRIME DP - Unbound Medicine ER -